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Improving the secretory expression of active recombinant AaIT in Pichia pastoris by changing the expression strain and plasmid

  • Hongbo LiEmail author
  • Yuxian XiaEmail author
Original Paper

Abstract

Scorpion long-chain insect selective neurotoxin AaIT has the potential to be used against agricultural insect pests. However, there is still a lack of a heterologous gene expression system that can express AaIT efficiently. Here, using X33 as the host strain and pPICZαA as the expression vector, one transformant had the highest expression of recombinant AaIT (rAaIT) was obtained, and secreted as high as 240 mg/l rAaIT in fed-batch fermentation. Secretory rAaIT was purified by Ni2+-nitriloacetic affinity and CM chromatography, and 8 mg of high purity rAaIT were purified from 200 ml fed-batch fermentation cultures. Injecting silkworm (Bombyx mori Linnaeus) and Galleria mellonella larvae with rAaIT resulted in obvious neurotoxin symptoms and led to death. These results demonstrate that a large amount of anti-insect active rAaIT could be prepared efficiently.

Keywords

Scorpion insect neurotoxin AaIT Pichia pastoriFed-batch Expression and purification 

Notes

Acknowledgements

This work was supported in part by Funds from Project funded by China Postdoctoral Science Foundation (2016M590867), Project funded of Chongqing Postdoctoral Science Foundation (Xm2016075) and Scientific Research Fund of Hunan Provincial Education Department (15A147).

Supplementary material

Supplementary material 1 (MOV 25563 KB)

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Copyright information

© Springer Nature B.V. 2018

Authors and Affiliations

  1. 1.Postdoctoral Mobile Station of Biology, Genetic Engineering Research Center, College of Life SciencesChongqing UniversityChongqingChina
  2. 2.The Key Laboratory of Research and Utilization of Ethnomedicinal Plant Resources of Hunan Province, College of Biological and Food EngineeringHuaihua UniversityHuaihuaChina

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